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Identification of vitamin A-free cells in a stellate cell-enriched fraction of normal rat liver as myofibroblasts

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Abstract

Myofibroblasts (MFs) as well as hepatic stellate cells (HSCs) are known to be involved in liver fibrogenesis. Quiescent HSCs (qHSCs) in culture have been thought to differentiate to replicative activated HSCs (aHSCs). In this study a qHSC-enriched fraction isolated by Nycodenz-isodensity centrifugation was separated with a fluorescence-activated cell sorter, which revealed the presence of a small fraction (occupancy rate = 0.4%) of cells that did not show vitamin A-autofluorescence under ultraviolet (UV)-irradiation (UV− cells). The remaining vitamin A-containing cells were autofluorescent (UV+) and originally expressed markers of qHSCs, and, in culture, did not grow, lost vitamin A, and expressed markers of aHSCs. UV− cells showed morphology of MFs, and, in culture, grew to form colonies and expressed markers of MFs. These results indicated that UV+ and UV− cells represent qHSCs and MFs, respectively, and that aHSCs have no growth potential and are a cell-type distinct from proliferative MFs. Gene expression profiles of UV− cells (MFs) newly identified gremlin as one of MF-preferential genes and its proteins were localized around fibrotic septa in rat and human livers. In addition, we suggested that the qHSC-enriched fraction included approximately 6% of liver MFs.

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Acknowledgments

We thank Drs. R. Timpl and T. Sasaki at the Max-Planck Institute for kindly providing us with the antisera against mouse fibulin-2, and C. Ohnishi for an excellent FACS operation.

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Authors and Affiliations

  1. Developmental Biology Laboratory, Department of Biological Science, Graduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashihiroshima, Hiroshima, 739-8526, Japan

    Tomohiro Ogawa, Masanobu Obara & Katsutoshi Yoshizato

  2. Hiroshima University 21st Century COE Program for Advanced Radiation Casualty Medicine, Developmental Biology Laboratory, Department of Biological Science, Graduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashihiroshima, Hiroshima, 739-8526, Japan

    Tomohiro Ogawa & Katsutoshi Yoshizato

  3. Yoshizato Project, Cooperative Link of Unique Science and Technology for Economy Revitalization, Hiroshima Prefectural Institute of Industrial Science and Technology, 3-10-32 Kagamiyama, Higashihiroshima, Hiroshima, 739-0046, Japan

    Tomohiro Ogawa, Chise Tateno & Katsutoshi Yoshizato

  4. Hiroshima Tissue Regeneration Project, Hiroshima Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence, Japan Science and Technology Organization, Hiroshima Prefectural Institute of Industrial Science and Technology, 3-10-32 Kagamiyama, Higashihiroshima, Hiroshima, 739-0046, Japan

    Tomohiro Ogawa, Chise Tateno, Kinji Asahina & Katsutoshi Yoshizato

  5. Liver Project Research Center, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan

    Chise Tateno & Katsutoshi Yoshizato

  6. Department of Pathological Biochemistry, Medical Research Institute, Tokyo Medical and Dental University, 2-3-10 Kandasurugadai, Chiyoda-ku, Tokyo, 101-0062, Japan

    Kinji Asahina

  7. Department of Hepatology, Graduate School of Medicine, Osaka City University, 1-4-3 Asahi-Machi, Abeno-ku, Osaka, 545-8585, Japan

    Hideki Fujii & Norifumi Kawada

Authors
  1. Tomohiro Ogawa
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  2. Chise Tateno
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  3. Kinji Asahina
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  4. Hideki Fujii
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  5. Norifumi Kawada
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  6. Masanobu Obara
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  7. Katsutoshi Yoshizato
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Corresponding author

Correspondence to Katsutoshi Yoshizato.

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Ogawa, T., Tateno, C., Asahina, K. et al. Identification of vitamin A-free cells in a stellate cell-enriched fraction of normal rat liver as myofibroblasts. Histochem Cell Biol 127, 161–174 (2007). https://doi.org/10.1007/s00418-006-0237-7

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  • DOI: https://doi.org/10.1007/s00418-006-0237-7

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